HES1 CRISPR Activation Plasmid (h): sc-400387-ACT

HES1 (hairy and enhancer of split 1) encodes a basic helix-loop-helix transcriptional repressor that functions as a core effector of Notch signaling to maintain progenitor states and suppress lineage-specific differentiation programs. By recruiting corepressor complexes and modulating transcriptional oscillations, HES1 influences cell cycle progression, neural and hematopoietic fate decisions, and stem cell maintenance, and it integrates inputs from pathways such as Wnt and Hedgehog. Dysregulated HES1 activity has been implicated in aberrant developmental programs and oncogenic transcriptional networks, including contexts where Notch pathway alterations contribute to uncontrolled proliferation. As a nodal regulator of differentiation and plasticity, HES1 is frequently studied in models of tumor biology, neurodevelopment, and regenerative processes.

HES1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HES1 expression without altering the underlying DNA sequence.

HES1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HES1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the HES1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HES1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HES1 locus and enabling the study of HES1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HES1 pathway restoration in tumor cells with silenced or reduced HES1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.